Erik A. Burton

Inhibition of Escherichia coli Biofilm Formation by Self-Assembled Monolayers of Functional Alkanethiols on Gold

ABSTRACT Bacterial biofilms cause serious problems, such as antibiotic resistance and medical device-related infections. To further understand bacterium-surface interactions and to develop efficient control strategies, self-assembled monolayers (SAMs) of alkanethiols presenting different functional groups on gold films were analyzed to determine their resistance to biofilm formation. Escherichia coli was labeled with green florescence protein, and its biofilm formation on SAM-modified surfaces was monitored by confocal laser scanning microscopy. The three-dimensional structures of biofilms were analyzed with the COMSTAT software to obtain information about biofilm thickness and surface coverage. SAMs presenting methyl, l-gulonamide (a sugar alcohol tethered with an amide bond), and tri(ethylene glycol) (TEG) groups were tested. Among these, the TEG-terminated SAM was the most resistant to E. coli biofilm formation; e.g., it repressed biofilm formation by E. coli DH5α by 99.5% ± 0.1% for 1 day compared to the biofilm formation on a bare gold surface. When surfaces were patterned with regions consisting of methyl-terminated SAMs surrounded by TEG-terminated SAMs, E. coli formed biofilms only on methyl-terminated patterns. Addition of TEG as a free molecule to growth medium at concentrations of 0.1 and 1.0% also inhibited biofilm formation, while TEG at concentrations up to 1.5% did not have any noticeable effects on cell growth. The results of this study suggest that the reduction in biofilm formation on surfaces modified with TEG-terminated SAMs is a result of multiple factors, including the solvent structure at the interface, the chemorepellent nature of TEG, and the inhibitory effect of TEG on cell motility.

Molecular Gradients of Bioinertness Reveal a Mechanistic Difference between Mammalian Cell Adhesion and Bacterial Biofilm Formation

Chemical gradients play an important role in guiding the activities of both eukaryotic and prokaryotic cells. Here, we used molecularly well-defined chemical gradients formed by self-assembled monolayers (SAMs) on gold films to reveal that mammalian cell adhesion and bacterial biofilm formation respond differently to a gradient of surface chemistry that resists cell attachment. Gradient self-assembled monolayers (SAMs) consisting of two mixed alkanethiols were fabricated by differential exposure of the gold film to one alkanethiol, followed by soaking in another alkanethiol solution. A gradient in bioinertness that resisted cell attachment was created on SAMs from a gradient in the surface density of HS(CH2)11(OCH2CH2)3OH, backfilled with either HS(CH2)11OH or HS(CH2)11CH3. Measurements of the amounts of mammalian cells and bacterial biofilms on these gradient surfaces reveal that, for mammalian cells, a critical density of adhesion ligands from absorbed proteins on surfaces exists for supporting maximum adhesion and proliferation, whereas for the bacterium Escherichia coli , the amount of biofilm formed on surfaces increased linearly with the surface density of adhesive groups (methyl or hydroxyl groups) in different media. These results are consistent with mammalian cell adhesion requiring an anchorage via specific molecular recognitions and suggest that biofilms can form by immobilization of bacteria via nonspecific interaction between bacteria and surfaces.

Prolonged control of patterned biofilm formation by bio-inert surface chemistry.

A bio-inert surface chemistry was developed that can confine biofilm formation in designed patterns for at least 26 days.